Method for control of white muscle disease

ABSTRACT

White muscle disease is controlled by administering the combination of a selenium compound and vitamin E, by parenteral injection to any mammal or by oral ingestion in mammals other than herbivores.

United States Patent [191 Burns et al.

[ Aug. 13, 1974 METHOD FOR CONTROL OF WHITE MUSCLE DISEASE [73]Assignee: Chromalloy Pharmaceutical, lnc.,

Wilmington, Del.

[22] Filed: Apr. 28, 1967 [21] Appl. No.: 639,923

Related US. Application Data [63] Continuation-in-part of Ser. No.352,681, March 17,

1964, abandoned.

[52] US. Cl. 424/131, 424/284 [51] Int. Cl A61k 15/10 [58] Field ofSearch 424/284, 131

[56] References Cited OTHER PUBLICATIONS 8th Annual H. C. Burns Co.,Symposium, Selenium-Tocopheryl Deficiency Diseases, Jan. 27, 1963, pages4 and 5.

Trademark 764,570, BO-SE Feb. 11, I964, Dispensary of the US. publishedby J. B. Lipponcott Co., Philadelphia, 1955, page 1839.

Kendall, The California Veterinarian, Vol. 14, No. 1, page 39,Sept.-Oct. 1960.

Maplesden et al.,.l. Dairy Science, Vol. 43, pages 645-653, May 1960.

Modem Veterinary Practice, Sept. 1964, pages 59-63.

.Nesheim et al., J. Nutrition, Vol. 65, pages Scott et al., PoultryScience." Vol. 36, page 1155, 1957. Schwarz et al., J. Biol. Chem, Vol.233, pages 245-251, 1958.

Sharman, Veterinarian Record, Aug. 15, 1959, page 536.

Vet. Drug. Encyclopedia, 6th Ed, 1958, page 17. Welch et al., J. AnimalScience, Vol. 19, pages 620-628.

Zalkin et al., Arch. Biochem. Biophys, Vol. 91, pages 117-122, 1960.

Primary Examiner-Richard L. Huff Attorney, Agent, or F irm-CurtisiMorris& Safiordg Y Garv A. Parto yan, Esq.

[5 7] ABSTRACT 7 Claims, No Drawings METHOD FOR CONTROL OF WHITE MUSCLEDISEASE This is a continuation-in-part of our application Ser. No.352,681, filed Mar. 17, 1964, which was a continuation-in-part of ourapplications Ser. No. 300,684, filed Aug. 7, 1963, and Ser. No. 96,733,filed Mar. 20, 1961 all now abandoned.

The invention relates to a pharmaceutical composition and a process forcontrolling, i.e., preventing and treating, white muscle disease (hereincalled WMD) in mammals, including beasts and humans.

WMD in mammals is a clinical manifestation of a disease complex orsyndrome characterized by degenerative lesions in the musculature,notably hyalinization and calcium deposits in cardiac and skeletalmuscles, including those of the diaphragm, intercostal and limb muscles,genito-urinary, digestive, neurological and skin connective tissue, etc.Early signs are generally weakness and incoordination, although theaffected mammals may die at birth. Weakness may progress until an animalcannot rise. Breathing may be labored (dyspnea) and cardiacinsufficiency may result in death from asphyxia accompanied by edema andcongestion of the lungs. When the heart muscle is affected (myocarditis)the mammal, upon exertion, may drop dead (cardiac arrest). Diarrhea orscours (enteritis) and unthriftiness (ill-thrift) are often observed. Ahigh incidence of reproduction disturbances (barrenness; poor conceptionrate; abortions; still born offspring; premature births) may occur.Tongue paralysis (glossoplegia) and consequent inability to nurse hasoften been reported as a clinical sign of this syndrome.

This syndrome may be mild to quite striking in its early appearances.Losses of animals by death range from a few to a high percentage, up to100 percent, of the young animals. The economic consequences are severeand their magnitude is apparent from reports regarding Klamath County,Oregon, by the County Extension Agents, a cooperative including OregonState College and the United States Department of Agriculture. Thesereports show the annual production in the county of 40,000 calves and50,000 lambs annually, and losses from WMD by death and other causes wasconservatively estimated to be.$250,000.00 per year. The Farm Advisor,Cooperative Extension Work, University of California and the US.Department of Agriculture, Siskiyou and Modoc Counties California,cooperating, reported a conservative estimate of about 1,000 lambs lostyearly from WMD and that during the last year prior to the use of theproduct of this invention, some sheep men were reporting 40 percent losson new lambs due to this disease. The County Extension Agent,Cooperative Extension work, Oregon State College, the United StatesDepartment of Agriculture, and Union County, Oregon, cooperating,reported an estimate of between 20,000 and 30,000 calves and lambs lostduring the year 1960 due to WMD.

It is believed that WMD is caused by an impairment of activity ofselenium or vitamin E in the body, due to a failure of the body totransport or utilize these factors even though they may be present inthe diet. While occasional and unpredictable beneficial results had beenachieved by the administration of each of these factors separately, theresults were so inconsistent and unsatisfactory that no dependabletreatment was devised, and great economic loss continued prior to thepresent invention. It was generally held by the scientific communitythat selenium was an alternative to vitamin E as a prophylactic andtherapeutic agent in combating WMD, but selenium was considered to behazardous and therefore not administered extensively.

It is an object of this invention to provide a composition and a processfor the control of WMD which is highly dependable and effective, andwhich can be readily and precisely administered and is therefore safe bystandard procedures.

Another object is to provide a composition of the character describedwhich can be compounded at a reasonable cost, is stable, and retains itseffectiveness over a prolonged period of time.

Now in accordance with the invention, WMD in mammals is controlled bythe simultaneous administration of vitamin E and a selenium compoundunder conditions which avoid interference with utilization by the bodyby extraneous substances which may be present in the diet and inhibitutilization. The invention is based on the double discovery that (l) thesimultaneous administration of these factors produces a synergisticeffect and (2) that administration must be so effected as to avoidinterference by such extraneous inhibiting substances.

More particularly, it was found that interference by such extraneoussubstances can be effected by parenteral, e.g., subcutaneous orintramuscular, injection of an injectable composition consistingessentially of vitamin E and a selenium compound. Such extraneoussubstances are often present in the feed of herbivorous animals, andexamples are compounds that contain sulfur, phosphorus and nitrates. Onthe other hand, nonherbivores, i.e., carnivores and omnivores, do notusually include large amounts of such substances in their diet, and thetwo synergistically acting materials can be administered orally. In thiscase, however, the synergism is less than optimum, although ademonstrable and satisfactory clinical response is achieved. Further, itis in this case preferable to encapsulate the vitamin E and seleniumcompound, both for the purpose of controlling the dose administered andinsuring the simultaneous entry of these factors into the body of thenon-herbivore.

The present inventors postulated that the biochemical basis of thedegenerative disease condition in WMD rests in part on the impropermetabolism of lipids in the body as a result of an impairment orderangement of selenium and vitamin E activity. Both of these actprimarily as anti-oxidants in alternative pathways of intermediatemetabolism, preventing the formation of lipid peroxides and acting asfree-radical traps in neutraliz ing these damaging peroxides. Whenselenium and vitamin B are not available in cellular metabolism inadequate amounts or proportions, the resulting lipid peroxides causesevere cellular damage to non-selective tissues. This cellular necrosisproduces the degenerative changes ultimately leading to clinicalmanifestations of the disease. However, it was known that prior attemptsto control WMD by treating the disease as a dietary deficiency gave onlyundependable and unpredictable results. For these reasons no effectiveremedy was known. Further, there appeared to be no merit in a mixture ofmaterials for controlling WMD because, when both vitamin E and aselenium compound were included in the feed to chicken for the controlof exudative diathesis, vitamin E administered together with theselenium compound yielded no better results than the selenium compoundalone. (Nesheim and Scott, Journal of Nutrition, vol. 65, 1958, pp. 601618.)

The inventors further hypothesized that oral administration of seleniumand of vitamin E gave inconsistent and inconclusive results in herbivorabecause:

1. Inhibiting substances common to the diet of her bivora ofteninterfere with the adsorption, transport and utilization of thesefactors. It is necessary to avoid the presence of such inhibitingsubstances.

2. ln non-herbivorous mammals these inhibiting substances areconsiderably less and often occur in such small amounts as not torequire the precaution noted in (l).

3. There exists a variation in the ability of animals to adsorb,transport and utilize selenium and/or vitamin E in their respectivebodies.

' 4. Both of these factors must be present. It was hypothesized thatselenium plays a necessary part in the transport of vitamin E across thecell membrane wall.

Regardless of the correctness of the above-stated theories, use ofmixtures of vitamin E and a selenium compound unexpectedly led todependable and eminently satisfactory results in the treatment andprevention of WMD in mammals. In the course of extensive tests, it wasfound that these factors act synergistically. One possible explanationis that an adequate supply of both factors at the cellular level isnecessary to assure dependable efficiency in the utilization of thesesubstances by the animal. However, the true cause of this synergism isnot known.

Synergism is demonstrated by tests which demonstrated an enhancement ofpotency of selenium and vitamin E by two to six times when they areadministered simultaneously and the foregoing precaution againstinhibiting substances is taken. Specifically, clinical evaluations showthat selenium and vitamin E, administered simultaneously, produce aneffectiveness of over 99 per cent, both therapeutically and as apreventative; this is in contrast to about 44 to 48 per centeffectiveness for selenium alone and about 12 to 16 per cent for vitaminE alone. These tests were performed on herds of lambs and calvesproduced by stock grazing within a basin of generally uniform conditionsof climate and forage properties, known to contain sufficient vitamin Eto meet the normal requirements of the animals.

The effectiveness of the mixture has been established by the highlysuccessful results achieved in the administration of over five millioninjected doses, most of which were administered prophylactically. Theseresulted in complete control of WMD. Thus, when an animal shows theslightest symptoms of WMD and is treated, recovery results is betterthan 99 per cent of the animals. The treatment has also been appliedprophylactically, with almost complete elimination of WMD. In one areain northern California comprising four counties, it was estimated by theDepartment of Agriculture officials of California that annual savings offrom one-half to one million dollars have been effected by the livestockindustry through application of the invention. Savings are now estimatedby Oregon State University to be ten million dollars annually for thePacific Northwest.

Experimental work with the preparation of the invention has been carriedout by many doctors of veterinary medicine and other scientificinvestigators, including as representative of the group: Gordon Shultz,D.V.M., Department of Agriculture, State of California; B. D. Kuhl,D.V.M., of Baker, Oregon; C. C. Beck, D.V.M., Department of Surgery andMedicine, Michigan State University; Jack A. Tucker, D.V.M., of MountShasta, California; Don L. Mace, D.V.M., M.P.H., California StateDepartment of Agriculture, Bureau of Animal Health; 0. K. Kendall,D.V.M., of Yreka, California; Edward M. Smith, D.V.M., of Orland,California; R. M. Stauffer, D.V.M., of Red Bluff, California; H. M.Adams, D.V.M., of Astoria, Oregon; H. E. Hill, D.V.M., of ImperialBeach, California; James M. Harris, D.V.M., of Oakland, California; MaxW. Colton, D.V.M., of Santa Barbara, California; Harry A. Leonard,D.V.M., of Klamath Falls, Oregon; Jack L. Kohler, D.V.M., of Oakland,California; M. L. Miller, D.V.M., of Elma, Washington; A. L. Tappel, Ph.D., Professor, Department of Food and Science Technology, University ofCalifornia; Fred J. Wedam, D.V.M., of Klamath Falls, Oregon; JosephRiker, D.V.M., of Klamath Falls, Oregon; and George Crenshaw, D.V.M., ofOrland, California. Findings in such experimental work have demonstratedthe effectiveness of the present medication in the treatment and controlof the group of metabolic degenerative diseases herein above discussed.

Various vitamin E and selenium compounds and derivatives have beenevaluated, and in each case the lowest common denominator is theselenium atom or the tocopherol fraction we recognize as vitamin E.Therefore, selenium and vitamin E are the basic biochemically activefactors responsible for the clinical effects observed and they eachmight be employed in various forms and compounds designed to introduceproperly and effectively these basic factors, (selenium and vitamin E)into the body and thereby exert its pharmacodynamic effects uponmetabolism. Examples of selenium compounds which may be used includesodium selenate, barium selenate, selenomethionine, di-seleno-divalericacid, sodium selenite, potassium selenite, barium selenite,seleno-cystene, selenium oxide, potassium selenocynote, etc.

Examples of vitamin E which may be used include alpha tocopherol, betatocopherol, gamma tocopherol, delta tocopherol, epsilon tocopherol, zetatocopherol, eta tocopherol, d-alpha tocopherol, d-l-alpha tocopherol,d-l-alpha tocopheryl acetate, mixed natural tocopherols, disodium saltof d-l-alpha tocopherol, alphatocopheryl succinate, and the tocopherolgroup individually and collectively.

It is, at present, preferred to use selenium in the form of sodiumselenite, because of its solubility, relatively low toxicity, andbio-potency. Also, at present it is preferred to use vitamin E in theform of a member of the class consisting of d-alpha tocopheryl acetateor alpha tocopheryl succinat e because of the absence of side effects,greater bio-potency and the ease with which they can be combined withthe selenium compound to produce a safe and effective solution forinjection or encapsulation. However, the biochemical action and effectsas above discussed reside in the basic factors, selenium and vitamin E,and not in the vehicular compound of each employed to carry them intothe body.

Several examples are presented of the preparation of the pharmaceuticalcomposition of this invention. The

first five are for parenteral injection and the sixth for oralingestion. Generally, sodium selenite is included within a range ofabout 0.55 mg. to 11 mg. per cubic centimeter, and d-alpha tocopherylacetate may be included in a range of about to 100 mg. per cubiccentimeter.

EXAMPLE 1: Each c.c. contains:

Selenium (as sodium selenite,

0.55 mgm. 0.25 mg Vitamin E (as cl-alpha-tocopheryl acetate. 50 mgm) 68[.U.*

Polysorbate 8O U.S.P. 250 mg Thimerosal N.F. 1 mg Water q.s. to 1 c.c.

EXAMPLE ll: Each c. c. contains:

Selenium (as sodium selenite, 1.00 mg. 2.2. mg)

Vitamin E (as d-alpha-tocopheryl acetate, 50 mgm) 68 l.l J.*

Polysorbate 80 U.S.P. 250 mg.

Thimerosal NF. 1 mg.

Water q.s. to l c.c.

EXAMPLE Ill: Each c.c. contains:

Selenium (as sodium selenite, ll mg) 5.0 mg.

Vitamin E. (as d-alpha-tocopheryl acetate, 50 mgm) 68 l.U.*

Polysorbate 80 U.S.P. 250 mg Thimerosal N.F. 1 mg Water q.s. to l c.c.

EXAMPLE IV: Each c. c. contains:

Selenium (as sodium selenite, 2.5 mg. 5.5 mg

Vitamin E (as d-alpha-tocopheryl acetate, 50 mg) 68 1.U.

Polysorbate 80 U.S.P. 10 mg.

Propylparaben 1 mg.

Water for injection q.s. to l c.c.

EXAMPLE V: Each c.c. contains:

Selenium (as sodium selenite, 1.0 mg. 2.2 mg) Vitamin E (asd-alpha-tocopheryl acetate, 50 mg) 68 l.U.*

Polysorbate 80 U.S.P. 10 mg.

Propylparaben 1 mg.

Water for injection q.s. to l c.c. EXAMPLE V1: (for ingestion) Eachcapsule to contain:

Vitamin E 68 l.U.* (From d-alpha tocopheryl acid succinate containing1210 [.U. per gm. 59 mg.

Selenium 1 mg (From sodium selenite 2.3 mg.)

Magnesium stearate 25 mg (as filler and lubricant) International UnitsSodium selenite (Na SeO contains 45.67 per cent selenium. It is a whitecrystalline powder. lts crystals are tetragonal prisms, stable in air.It is water soluble and is alkaline to litmus paper.

D-alpha tocopheryl acetate N.F. is a light brownish, nearly odorless,clear viscous oil having a melting point at approximately 25C. It isinsoluble in water and has a specific gravity of 0.950 to 0.964inclusive.

Polysorbate 8O U.S.P. is a lemon to amber-colored oily liquid having afaint characteristic odor, and a warm somewhat bitter taste. It has aspecific gravity of between 1.06 and 1.1. The pH of a l in 20 solutionis between 6 and 8. Its principal use in the present composition is as asuspending or emulsifying agent for the d-alpha tocopheryl acetate. Itsexternal phase is water. Polysorbate is produced by Atlas Powder Co.under the trademark TWEEN.

Thimerosal N.F. is a light, cream-colored crystalline powder with slightcharacteristic odor. It is soluble in water and a 1 in 100 solution hasa pH of 6.7 Thimerosal is produced by Mann Fine Chemical Co. of NewYork, NY. and by Eli Lilly Co. of Indianapolis, Indiana, under thetrademark MERTHIOLATE. It functions in the present preparation as abacteriostatic agent.

Propylparaben occurs as small, colorless crystals, or as a whitecrystalline powder. It has a melting point of about 98 C. One gramdissolves in 2000 mls. of water.

D-alpha tocopheryl acid succinate occurs as a white crystalline powder.It has little or no taste or odor. It is stable to air, but is unstableto alkali and to heat. The absorptivity of d-alpha tocopheryl acidsuccinate, determined in alcohol at 284 mu, is not less than 35 and notmore than 40. One gm. of d-alpha tocopheryl acid succinate requires forneutralization not less than 18.0 ml and not more than 19.3 ml ofO.lN-NaOl-l. Melting Rangebetween 73 and 78 C.

Magnesium stearate occurs as a fine white, bulky powder having a slightodor. It is insoluble in l-IOH, in alcohol and in ether.

The preparations, Examples I, II and Ill, may be compounded in thefollowing manner. The D-alpha tocopheryl acetate and the Polysorbate 80are preferably heated before admixing is attempted. A temperature ofapproximately 60 C. is satisfactory. The Polysorbate 80 is added to thed-alpha-tocopheryl acetate with constant agitation which is preferablyquite vigorous. The sodium selenite and thimerosal are diluted in watersolutions and preferably for this purpose about one-half of the requiredwater, preferably freshly distilled, is heated to about 60 C. and thesodium selenite and thimerosal stirred into this water in quantity untilsolution is effected. The water solution is then added slowly to thefirst mentioned solution with constant agitation and the mixture is thenbrought to final volume with additional distilled water with agitation.The product is allowed to cool and then sterilized by suitablefiltration through a Micro porcelain filter and vial aseptically. Theproduct is opaque until it is cool.

Preparations, Examples IV and V, may be compounded in the followingmanner, the example here given being for 45000 mls batch:

Dissolve 45 gms of propylparaben in 250 gms of polysorbate with the aidof heat. Add 40 liters of water. Dissolve 247.59 gms of sodium selenite,adjust pH to 6.5 7.5 with HCl, and filter into calibrated bottle througha Millipore using an HA membrane. Heat to 60 in autoclave.

Using d-alpha tocopheryl acetate having 1360 U/mg, weigh out 2475 gmsand add 245 gms of polysorbate into a beaker. Heat to 60 C. Add to thehot parabenselenium-polysorbate solution. Stir well, cool, adjust tofinal volume, and check pH. Fill into vials with emulsion under constantvigorous agitation. Sterilize by autoclaving at 240250 F. for minutesShake well about 1 hour after removing from semi-colon pH tolerance: 5.57.5.

The preparation of Example V1 is as follows:

Triturate the selenium into the vitamin E. Mix in the magnesium stearatevery thoroughly and pass through a 14 mesh screen several times.

Run at 86.3 mg. per capsule (running tolerance: 80 to 93 mg.) on a Lillycapsulating machine, using No. 3 yellow-green hard shell capsules.

We have found the products produced as above to be completely stable.Stability tests performed 3 months after manufacture have indicated nodeterioration.

The minimum recommended dose of selenium is about one-half milligram per100 pounds of body weight, and the maximum dosage would be about 10milligrams per 100 pounds of body weight. The minimum recommended dosagefor vitamin E would be about International Units per 100 pounds of bodyweight and the maximum dose would be about 1500 International Units per100 pounds of body weight.

The following Table is furnished as a guide for proper treatment of theanimals noted:

PREPA RATION EXAMPLE USED Newborn Lambs Lambs-2 weeks old Lambs-2 weeksold Newborn calves Pregnant Ewes Weaner Calves Pregnant Cows Horses DogsSwine Cats Chickens Turkeys Antelope Kangaroo Dogs and Cats Human Swinel c.c.

l c.c.

2% c.c. per 100 lbs. I c.c. per 200 lbs.

I c.c. per 200 lbs.

1 c.c. per 100 lbs.

1 c.c. per 20 lbs. 2% c.c. per 100 lbs. I c.c.

l c.c.

l c.c.

2% c.c. per 100 lbs. 2% c.c. per 100 lbs.

I capsule per lbs. daily to weekly 1 capsule per 40 lbs. daily to weeklyl capsule per 40 lbs. daily to weekly body weight Best results have beenobtained with lambs by injec- October 23, 1963, 5.0 mg. selenium and 136l.U. d-alpha tocopheryl acetate October 29, 1963, 5.0 mg. selenium and136 LU. d-alpha tocopheryl acetate November 5, 1963, 5.0 mg. seleniumand I36 l.U. d-alpha tocopheryl acetate November 13, 1963, 2.5 mg.selenium and 68 LU. d-alpha tocopheryl acetate November 26, 1963, 5.0mg. selenium and 136 l.U. d-alpha tocopheryl acetate RESULTS: 5 daysafter initial dose partial relief was obtained followed by gradual andcontinuous improvement until complete absence of pain on all movementsexcept laterally up to level of shoulder after 3 weekly doses. Lateralmovement greatly improved but not asymptomac.

PATIENT: Dr. F. G. Rankin SEX: Male ADDRESS: Salem, Oregon AGE: 50

DOCTOR: Same DATE: 1 l/4/63 ADDRESS Salem, Oregon WEIGHT: 250 lbs.

HISTORY: Chronic and persistent rheumatic-arthritic neuralgia insacro-iliac region gradually increasing in intensity over a 25 yearperiod. Sleep interrupted by pain permitting only 1% to 2 hours at atime. A sacroiliac belt (14 inches) has been worn for 7 years.DIAGNOSIS: Chronic progressive rheumatic-arthritic neuralgia in spine(sacro-iliac).

TREATMENT: A combination S-T product (Burns) containing 2-% mg. seleniumand 170 I.U. vitamin E per dose, 2 times weekly commencing August 25,1963, to date.

RESULTS: 48 hours after initial dose, relief from pain alloweduninterrupted sleep for 4% hours consecutively and could function forhours without sacro-iliac belt during waking period. Improvement gradualand steady to present so that uninterrupted sleep period is now 6 hours.

PATIENT: Mrs. Rankin SEX: Female ADDRESS: 2360 Fairgrounds Road AGE: 50

Salem, Oregon DATE: 11/4/63 DOCTOR: Dr. F. G. Rankin WEIGHT: lbs.

Salem, Oregon HISTORY: Intermittent muscular spasms for over 3 yearswith extreme pain interfering with sleep and rest. Tranquilizers andnarcotics analgesics offering some temporary relief.

DIAGNOSIS: Spastic Neuro-Myositis.

TREATMENT: Burns -S-T product containing per dose: 2- /2 mg. seleniumand 170 I.U. vitamin E per week to present (4 weeks).

RESULTS: Within 24 hours after initial dose, spasticity markedly reducedand at present after 4 weekly doses patient is asymptomatic; no pain, nospasm and no clinical evidence or original disorder.

PATIENT: Gwynne McKenzie SEX: Female ADDRESS: 5823 Trask AGE: 60

Oakland, California DOCTOR: Clinic Doctors DATE: 2/l4/64 ADDRESS: KaiserHospital WEIGHT: I30 lbs.

HISTORY: Very severe headaches persistent and chronic migrating painalong spinal column from between shoulders, up neck to head. Treatmentconsisted of Butazolidin, vitamins, Hormones and Corticosteroids withcontrol of pain initially and gradual diminishment in effectiveness.

DIAGNOSIS: Neuro-Muscular-Skeletal Degeneration.

TREATMENT: One Capsule, Example II above, every other day for 2 weekscommencing Nov. 7, 1963. Discontinued for 3 weeks. Resumed treatmentwith 1 Capsule every other day for 2 weeks, at end of this period dosagedecreased to l Capsule per week. All other medication was discontinuedat onset of treatment with Capsules, Example II.

RESULTS: Within 1 week of treatment with preparation of the presentinvention a slight and gradual reduction in pain and general discomfort.Within days of initial dose complete relief from symptoms was achieved.2 weeks after discontinuing Capsules a gradual return of pain occurredand Capsule treatment was resumed on 3rd week with prompt relief. OneCapsule/week has been maintaining patient asymptomatic.

We claim:

1. A method for controlling white muscle disease syndrome in mammalswhich comprises parenterally injecting into said mammals a medicallyeffective quantity of a product consisting essentially of a synergisticmixture of vitamin E and sodium selenite, said mixture being in apharmaceutically acceptable aqueous carrier and the weight ratio ofvitamin E to selenium in said product being in the range 10:1 to 200:1,said product having a maximum selenium content of about 1.2 percent byweight.

2. A method according to claim 1 and wherein the quantity of saidproduct administered and the composition thereof are such that about 25to 1500 LU. of vitamin E and about 0.5 to 10 milligrams of selenium per100 pounds of body weight of the mammal are administered.

3. A method for controlling white muscle disease syndrome in mammalswhich comprises parenterally injecting into said mammals a medicallyeffective quantity of a product consisting essentially of a synergisticmixture of vitamin E and sodium selenite in a pharmaceuticallyacceptable aqueous carrier, said product containing from about 10 to 100milligrams per cubic centimeter of vitamin E and about 0.55 to 11milligrams per cubic centimeter of sodium selenite.

4. Method of controlling white muscle disease in newborn and young lambsby injecting them according to claim 3 with approximately 1 cubiccentimeter of said product at birth followed by approximately 4 cubiccentimeters about 2 weeks later, said product containing about 50milligrams per cubic centimeter of vitamin E and about 0.55 milligramsper cubic centimeter of sodium selenite.

5. Method of controlling white muscle disease in newborn calves byinjecting them according to claim 4 with approximately 2 cubiccentimeters of said product, said product containing about 50 milligramsper cubic centimeter of vitamin E and about 2.2 milligrams per cubiccentimeter of sodium selenite.

6. Method of controlling white muscle disease in pregnant ewes whichcomprises injecting them according to claim 4 with approximately 2%cubic centimeters of said product per 100 lbs. of body weight, saidproduct containing about 50 milligrams per cubic centimeter of vitamin Eand about 2.2 milligrams per cubic centimeter of sodium selenite.

7. Method of controlling white muscle disease in weaner calves andpregnant cows which comprises injecting them according to claim 4 withapproximately the quantity of said product required to inject about 50milligrams of vitamin E and about ll milligrams of sodium selenite per200 lbs. of body weight.

UNITED STATES PATENT OFFICE 7 CERTIFICATE OF CORRECTION Patent No.3,829,566 Dated August 13, 1974 Inventor(s)' H y ns 'et a1 It iscertified that error appears in the'above-identified patent and thatsaid Letters'Patent are hereby corrected as shown below:

First Page, left column, line 4 from bottom, "U.S. should be UnitedStates of America,- and "Lipponcott" should be --Lippincott,; 7 rightcolumn, line 3, delete quote mark before "Sept. line 11, before"Veterinarian" insert -Theline 13, delete both quote marks and insertquote marks around Vet. Drug. Encyclopedia-, line 20, change "Garv" to-Garo; Col 1, line 8 after "1961" insert a comma, line 49, after"Counties" insert a comma; Col. 3, line 55, after "results" change "is"to -in; Col. 4, line 36, delete the comma, line 43, change "cynote" tocya'nate--, line 54, after "selenite" delete the comma, line 55, after"toxicity" delete the comma; Col. 5, line 12, change "0. 55 mgm." to -0.55 mg) lines 14, 23 and 33, in each instance change "50 mgm" to --50mg--, line 40 insert a close parenthesis after "5.5 mg" line 61, inserta close parenthesis after "59 mg. Col. 7, line 10, change "semi-colon"to -autoclave;--; Col. 8, line 28, change "asymptomac" to--asymptomatic--; Col. 9, line 7, change "or" to --of-; Col. 10, lines29, 36 and- 44 in each instance change "4" to 3-.

Signed and sealed this 14th day of January 1975.-

(SEAL) Attest:

GIBSON JR. 0. MARSHALL DANN fi g t i ng Officer Commissioner of PatentsFORM po'mso (1069) uscoMM-Dc 60378-P69 a U.S. GOVERNMENT PRINHNG OFFICEl9? 0- 66 334

2. A method according to claim 1 and wherein the quantity of saidproduct administered and the composition thereof are such that about 25to 1500 I.U. of vitamin E and about 0.5 to 10 milligrams of selenium per100 pounds of body weight of the mammal are administered.
 3. A methodfor controlling white muscle disease syndrome in mammals which comprisesparenterally injecting into said mammals a medically effective quantityof a product consisting essentially of a synergistic mixture of vitaminE and sodium selenite in a pharmaceutically acceptable aqueous carrier,said product containing from about 10 to 100 milligrams per cubiccentimeter of vitamin E and about 0.55 to 11 milligrams per cubiccentimeter of sodium selenite.
 4. Method of controlling white muscledisease in newborn and young lambs by injecting them according to claim3 with approximately 1 cubic centimeter of said product at birthfollowed by approximately 4 cubic centimeters about 2 weeks later, saidproduct containing about 50 milligrams per cubic centimeter of vitamin Eand about 0.55 milligrams per cubic centimeter of sodium selenite. 5.Method of controlling white muscle disease in newborn calves byinjecting them according to claim 4 with approximately 2 cubiccentimeters of said prOduct, said product containing about 50 milligramsper cubic centimeter of vitamin E and about 2.2 milligrams per cubiccentimeter of sodium selenite.
 6. Method of controlling white muscledisease in pregnant ewes which comprises injecting them according toclaim 4 with approximately 2 1/2 cubic centimeters of said product per100 lbs. of body weight, said product containing about 50 milligrams percubic centimeter of vitamin E and about 2.2 milligrams per cubiccentimeter of sodium selenite.
 7. Method of controlling white muscledisease in weaner calves and pregnant cows which comprises injectingthem according to claim 4 with approximately the quantity of saidproduct required to inject about 50 milligrams of vitamin E and about 11milligrams of sodium selenite per 200 lbs. of body weight.